The therapeutically anti-prion active antibody-fragment scFv-W226: paramagnetic relaxation-enhanced NMR spectroscopy aided structure elucidation of the paratope-epitope interface.
J Biomol Struct Dyn
; 28(1): 13-22, 2010 Aug.
Article
en En
| MEDLINE
| ID: mdl-20476792
ABSTRACT
Antibodies have become indispensable reagents with numerous applications in biological and biotechnical analysis, in diagnostics as well as in therapy. In all cases, selective interaction with an epitope is crucial and depends on the conformation of the paratope. While epitopes are routinely mapped at high throughput, methods revealing structural insights on a rather short timescale are rare. We here demonstrate paramagnetic relaxation-enhanced (PRE) NMR spectroscopy to be a powerful tool unraveling structural information about epitope-orientation in a groove spanned by the complementary determining regions. In particular, we utilize the spin label TOAC, which is fused to the peptidic epitope using standard solid-phase chemistry and which is characterized by a reduced mobility compared to, e.g., spin labels attached to the side-chain functionalities of cysteine or lysine residues. We apply the method to determine the orientation of helix 1 of the prion protein, which is the epitope for the therapeutically anti-prion active scF(v) fragment W226.
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Bases de datos:
MEDLINE
Métodos Terapéuticos y Terapias MTCI:
Terapias_mente_y_cuerpo
/
Relajacion
Asunto principal:
Conformación Proteica
/
Sitios de Unión de Anticuerpos
/
Priones
/
Espectroscopía de Resonancia Magnética
/
Anticuerpos de Cadena Única
/
Epítopos
Tipo de estudio:
Prognostic_studies
Idioma:
En
Revista:
J Biomol Struct Dyn
Año:
2010
Tipo del documento:
Article
País de afiliación:
Alemania